A simple one-step method for isolating highly purified plasmacytoid dendritic cells from human peripheral blood (78.33)

• Published in: The Journal of immunology (1950) 2009-04, Vol.182 (1_Supplement), p.78-78.33
• Main Authors: Kokaji, Andy I, Holland, Siobhan, Fairhurst, Maureen A, Thomas, Terry E, Guilbault, Benoit G
• Format: Article
• Language: English
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.182.Supp.78.33

Plasmacytoid dendritic cells (pDC) and myeloid DC (mDC) are the two major human dendritic cell populations. Each subset comprises only a small fraction of peripheral blood mononuclear cells (PBMC) and current protocols for their isolation are time consuming, involve multiple steps and often require special equipment. We have developed a rapid and efficient method for the isolation of pDCs from normal blood that yields excellent cell purities and recoveries. Briefly, PBMCs are isolated by Ficoll-Paque PLUS density gradient sedimentation and pDCs are isolated using immuno-magnetic, column-free, negative selection (EasySep®). Our EasySep® technology involves specifically labeling unwanted cells with a cocktail of bi-specific tetrameric antibody complexes and dextran-coated magnetic particles. Using a hand-held magnet, unwanted cells can then be easily removed from the unlabeled pDCs. Flow cytometric assessment of EasySep® isolated pDCs (Lin-, HLA-DR+, BDCA-4+) demonstrate purities of 93.8% ± 3.8% with cell recoveries of 65.6% ± 16.2% (n=9). Using our RoboSep® cell separator, the entire separation procedure can be fully automated with equivalent cell purities and recoveries. A.I.K is supported by an NSERC Industrial R&D Fellowship